Enhanced Ribozyme‐Catalyzed Recombination and Oligonucleotide Assembly in Peptide‐RNA Condensates

Abstract

The ability of RNA to catalyze RNA ligation is critical to its central role in many prebiotic model scenarios, in particular the copying of information during self‐replication. Prebiotically plausible ribozymes formed from short oligonucleotides can catalyze reversible RNA cleavage and ligation reactions, but harsh conditions or unusual scenarios are often required to promote folding and drive the reaction equilibrium towards ligation. Here, we demonstrate that ribozyme activity is greatly enhanced by charge‐mediated phase separation with poly‐L‐lysine, which shifts the reaction equilibrium from cleavage in solution to ligation in peptide‐RNA coaggregates and coacervates. This compartmentalization enables robust isothermal RNA assembly over a broad range of conditions, which can be leveraged to assemble long and complex RNAs from short fragments under mild conditions in the absence of exogenous activation chemistry, bridging the gap between pools of short oligomers and functional RNAs.

Peptide-based coacervates in therapeutic applications

“…Le Vay et al recently reported that in coacervation formed by poly- l -lysine (PLys) and a split HPz ribozyme, the ribozyme activity was increased significantly in different circumstances. The variant chain length of PLys showed that regardless of it being a longer peptide (PLys) 19–72 or a shorter peptide (PLys) 5–24 , a polycation peptide improved ribozyme activity ( Le Vay et al, 2021 ). This ability was suppressed at a high ratio of peptide, while at a low ratio, (PLys) 19–72 : RNA = 1.1:1 and (PLys) 5–24 : RNA = 2.3:1, the catalytic ability was strong.…”

Spatiotemporal Propagation of a Minimal Catalytic RNA Network in GUV Protocells by Temperature Cycling and Phase Separation

“…On the way towards reconstituting the cell origin, synthetic analogs are being constructed to mimic cell functions and structures [9] . Such synthetic organelles were shown to actively enrich biomolecules and efficiently promote enzymatic reactions [10–12] …”

Raumzeitliche Propagation eines minimalen katalytischen RNA‐Netzwerkes in GUV‐Protozellen durch Temperaturzyklen und Phasenseparation

“…Auf dem Weg hin zur Rekonstruktion des Zellursprungs werden synthetische Analoga genutzt, um Zellfunktionen und ‐strukturen nachzubilden [9] . Es wurde gezeigt, dass solche synthetischen Organellen dazu in der Lage sind, Biomoleküle anzureichern und enzymatische Reaktionen zu begünstigen [10–12] …”